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Practical aspects of implenting an antibody screening algorithm
Abstracts
S107
10.1 #139
PRACTICAL ASPECTS OF IMPLENTING AN ANTIBODY SCREENING ALGORITHM Lisa A. Geiselhart,1 Reva Goggins,1 Kristen Wolfe,1 Marc I. Lorber.1 1Surgery – Organ Transplantation & Immunology, Yale Univ. School of Medicine Box 208062, New Haven, CT Antibody screening has evolved from the simple microcytotoxicity assay into a complex array of methodologies including ELISA, flow cytometric & microsphere bead technologies. This multitude of screening technologies means that each lab has to develop their own algorithm to determine each sample PRA. The advantage of this algorithm is that it allows the technologist to determine the most appropriate screening method for each serum sample, while the disadvantage is that each sera has to be aliquoted differently. The chief complaint upon implementing new screening methods in our lab was that aliquoting was now too complicated, cumbersome, & inefficient, and involved too much thinking. In order to deal with this complex aliquoting scheme, we developed a series of mail merges that greatly facilitate this process. Our lab receives most of our monthly serum samples on the 1st Tues of the month. The samples are logged in & then merged with a Word label document. We print 1–7 labels per pt (depending upon the status & sensitization history) with name, SS#, ABO, organ, wait list status, unsens. or sens. (we plate these on separate cadaver donor trays), unt/HI, BCXM or not, screening instructions & serum date. This mail merge system can be used with any data source (xls, csv, mdb, dbf, etc). The Word “mail merge wizard” makes the process of setting up a merge document very easy, just go to “tools” “mail merge” to begin. Step 1 creates the label or form letter document. Step 2 opens your data source. Once the data source is linked to your document an “insert merge field” button appears, listing your data source fields, thus allowing you to chose the fields that you want to appear on your labels or worksheet. We use this “mail merge” concept to simplify a variety of tasks in our lab. Not only do we print our labels this way, but we also print our cadaver donor trays, our LAT–M ELISA tray worksheets, our Luminex tray worksheets, our request for serum letters, and our missing sample letters via mail merges. We want to share these templates & instructions with other labs because we believe that other labs might wish to utilize this mail merge scheme, which makes dealing with monthly pt serum samples much simpler & more efficient.
10.1 #140
COLLECTION AND ANALYSIS OF HLA TYPING DATA FROM MULTIPLE DNA–BASED TECHNOLOGIES USED TO TISSUE TYPE NMDP DONORS Mark A. Janzen,1 Alan Howard,1 Martin Maiers,1 Jon Sorbie,1 Michelle Setterholm,1 Carolyn K Hurley.2 1Scientific Services, National Marrow Donor Program, Minneapolis, Minnesota; 2Georgetown University Medical Center, Washington, DC The National Marrow Donor Program (NMDP) receives HLA typing results derived from a variety of DNA– based typing strategies. Collection and analysis of reagent specific typing data associated with reported results is used to validate submitted results and as new alleles are discovered in the future, to update stored HLA typings. In 2001, 238,000 newly recruited donors⬘ HLA typings were validated based on the stored sequences of the sequence specific oligonucleotide (SSO) reagents used by the NMDP laboratories, and the associated probe data stored for these donors. The next step was to collect reagent specific data derived from PCR with sequence specific primers (PCR–SSP), which is used as a supplemental typing strategy by the SSO laboratories to resolve ambiguous allele combinations. Moreover, PCR–SSP is the main testing methodology for the laboratories performing fast–turnaround time, high–resolution, HLA typing for the NMDP. A new database structure was designed to store the sequence information on each laboratory⬘s collection of SSO and SSP primer sequences (kit), and can accommodate an unlimited number or combination of SSOP and SSP reagents. A web– based HLA Typing Kit Registration & Maintenance Tool was developed to interface with the database and allows each laboratory⬘s kit to be updated as reagents are modified or added. Allele reactivity tables for the entered SSOP and SSP reagents can be generated and updated as new HLA Sequence Database versions are released. Initially, the tool will by used by NMDP staff for administering typing kits used by the network of contract laboratories, and in the future the laboratories will be able to directly register and update their own HLA typing kits using the tool. HLA–typings based on SSP reagent data now submitted to the NMDP are interpreted and validated in the same manner as HLA typings derived from SSO, and all SSP raw typing information is being stored for future reinterpretation. Finally, efforts are underway to also start incorporating data from sequence based typing methodologies.
S107
10.1 #139
PRACTICAL ASPECTS OF IMPLENTING AN ANTIBODY SCREENING ALGORITHM Lisa A. Geiselhart,1 Reva Goggins,1 Kristen Wolfe,1 Marc I. Lorber.1 1Surgery – Organ Transplantation & Immunology, Yale Univ. School of Medicine Box 208062, New Haven, CT Antibody screening has evolved from the simple microcytotoxicity assay into a complex array of methodologies including ELISA, flow cytometric & microsphere bead technologies. This multitude of screening technologies means that each lab has to develop their own algorithm to determine each sample PRA. The advantage of this algorithm is that it allows the technologist to determine the most appropriate screening method for each serum sample, while the disadvantage is that each sera has to be aliquoted differently. The chief complaint upon implementing new screening methods in our lab was that aliquoting was now too complicated, cumbersome, & inefficient, and involved too much thinking. In order to deal with this complex aliquoting scheme, we developed a series of mail merges that greatly facilitate this process. Our lab receives most of our monthly serum samples on the 1st Tues of the month. The samples are logged in & then merged with a Word label document. We print 1–7 labels per pt (depending upon the status & sensitization history) with name, SS#, ABO, organ, wait list status, unsens. or sens. (we plate these on separate cadaver donor trays), unt/HI, BCXM or not, screening instructions & serum date. This mail merge system can be used with any data source (xls, csv, mdb, dbf, etc). The Word “mail merge wizard” makes the process of setting up a merge document very easy, just go to “tools” “mail merge” to begin. Step 1 creates the label or form letter document. Step 2 opens your data source. Once the data source is linked to your document an “insert merge field” button appears, listing your data source fields, thus allowing you to chose the fields that you want to appear on your labels or worksheet. We use this “mail merge” concept to simplify a variety of tasks in our lab. Not only do we print our labels this way, but we also print our cadaver donor trays, our LAT–M ELISA tray worksheets, our Luminex tray worksheets, our request for serum letters, and our missing sample letters via mail merges. We want to share these templates & instructions with other labs because we believe that other labs might wish to utilize this mail merge scheme, which makes dealing with monthly pt serum samples much simpler & more efficient.
10.1 #140
COLLECTION AND ANALYSIS OF HLA TYPING DATA FROM MULTIPLE DNA–BASED TECHNOLOGIES USED TO TISSUE TYPE NMDP DONORS Mark A. Janzen,1 Alan Howard,1 Martin Maiers,1 Jon Sorbie,1 Michelle Setterholm,1 Carolyn K Hurley.2 1Scientific Services, National Marrow Donor Program, Minneapolis, Minnesota; 2Georgetown University Medical Center, Washington, DC The National Marrow Donor Program (NMDP) receives HLA typing results derived from a variety of DNA– based typing strategies. Collection and analysis of reagent specific typing data associated with reported results is used to validate submitted results and as new alleles are discovered in the future, to update stored HLA typings. In 2001, 238,000 newly recruited donors⬘ HLA typings were validated based on the stored sequences of the sequence specific oligonucleotide (SSO) reagents used by the NMDP laboratories, and the associated probe data stored for these donors. The next step was to collect reagent specific data derived from PCR with sequence specific primers (PCR–SSP), which is used as a supplemental typing strategy by the SSO laboratories to resolve ambiguous allele combinations. Moreover, PCR–SSP is the main testing methodology for the laboratories performing fast–turnaround time, high–resolution, HLA typing for the NMDP. A new database structure was designed to store the sequence information on each laboratory⬘s collection of SSO and SSP primer sequences (kit), and can accommodate an unlimited number or combination of SSOP and SSP reagents. A web– based HLA Typing Kit Registration & Maintenance Tool was developed to interface with the database and allows each laboratory⬘s kit to be updated as reagents are modified or added. Allele reactivity tables for the entered SSOP and SSP reagents can be generated and updated as new HLA Sequence Database versions are released. Initially, the tool will by used by NMDP staff for administering typing kits used by the network of contract laboratories, and in the future the laboratories will be able to directly register and update their own HLA typing kits using the tool. HLA–typings based on SSP reagent data now submitted to the NMDP are interpreted and validated in the same manner as HLA typings derived from SSO, and all SSP raw typing information is being stored for future reinterpretation. Finally, efforts are underway to also start incorporating data from sequence based typing methodologies.